Subpopulations of normal peripheral blood and bone marrow cells express a functional multidrug resistant phenotype.
Subpopulations of normal peripheral blood and bone marrow cells express a functional multidrug resistant phenotype.
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DOI：
10.1182/blood.v80.11.2729.bloodjournal80112729
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发表时间：
1992-12
期刊：
影响因子：
20.3
通讯作者：
D. Drach;Shourong Zhao;J. Drach;R. Mahadevia;Claus Gattringer;Heinz Huber;M. Andreeff
中科院分区：
文献类型：
--
作者：
D. Drach;Shourong Zhao;J. Drach;R. Mahadevia;Claus Gattringer;Heinz Huber;M. Andreeff
多重耐药基因MDR1在许多正常组织中表达，但对其在正常造血细胞中的表达知之甚少。使用单克隆抗体 C219 和流式细胞术分析，发现 P-糖蛋白 (P-gp) 在除粒细胞外的所有外周血 (PB) 亚群（CD4、CD8、CD14、CD19、CD56）中表达。为了特异性测定 MDR1 基因表达，通过荧光激活细胞分选 (FACS) 分离这些 PB 亚群，并通过聚合酶链式反应 (PCR) 分析 MDR1 mRNA。所有子集均通过 PCR 检测呈阳性，但在单核细胞和粒细胞中仅检测到极少量的 MDR1 mRNA。在 CD4+、CD8+、CD14+、CD19+ 和 CD56+ 细胞中检测到罗丹明-123 (Rh-123)（P-gp 功能的量度）的显着外流，但在粒细胞中未检测到。接下来，对 FACS 分选的骨髓 (BM) 细胞进行 PCR 分析，以评估不同成熟阶段的 MDR1 表达。前体细胞 (CD34+)、早期和晚期骨髓细胞 (CD33+/CD34+、CD33+/CD34-) 以及 B 细胞谱系的淋巴细胞 (CD19+/CD10+、CD19+/CD10-) 表达 MDR1 基因。 BM 单核细胞 (CD33++/CD34-) 呈阴性，在红系细胞 (血型糖蛋白 A+) 中检测到非常微弱的信号。在 CD34+、CD10+、CD33+ 和 CD33++ BM 细胞中发现显着的 Rh-123 外流，但在血型糖蛋白 A+ 细胞中没有发现。我们得出结论，PB 和 BM 淋巴细胞、PB 单核细胞、BM 祖细胞和未成熟骨髓细胞表达 MDR1 mRNA 和功能性 P-gp，但晚期 BM 单核细胞、红系细胞和 PB 粒细胞不表达。当在含有正常血细胞的肿瘤样本中测定 MDR1 表达时，必须考虑这些结果。
The multidrug-resistance gene, MDR1 is expressed in many normal tissues, but little is known about its expression in normal hematopoietic cells. Using the monoclonal antibody C219 and flow cytometric analysis, P-glycoprotein (P-gp) was found to be expressed in all peripheral blood (PB) subpopulations (CD4, CD8, CD14, CD19, CD56) except granulocytes. To specifically determine MDR1 gene expression, these PB subpopulations were isolated by fluorescence-activated cell sorting (FACS) and analyzed for MDR1 mRNA by polymerase chain reaction (PCR). All subsets were positive by PCR, but only minimal MDR1 mRNA was detected in monocytes and granulocytes. Significant efflux of Rhodamine-123 (Rh-123), a measure of P-gp function, was detected in CD4+, CD8+, CD14+, CD19+, and CD56+ cells but not in granulocytes. Next, PCR-analysis was performed on FACS-sorted bone marrow (BM) cells to assess MDR1 expression in different maturational stages. Precursors (CD34+), early and late myeloid cells (CD33+/CD34+, CD33+/CD34-) as well as lymphocytes of the B-cell lineage (CD19+/CD10+, CD19+/CD10-) expressed the MDR1 gene. BM monocytic cells (CD33++/CD34-) were negative, and a very weak signal was detected in erythroid cells (glycophorin A+). Significant Rh-123 efflux was found in CD34+, CD10+, CD33+, and CD33++ BM cells, but not in glycophorin A+ cells. We conclude that PB and BM lymphocytes, PB monocytes, BM progenitors, and immature myeloid cells, but not late BM monocytes, erythroid cells, and PB granulocytes, express MDR1 mRNA and a functional P-gp. These results have to be taken into account when MDR1 expression is determined in tumor samples containing normal blood cells.